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• W2017462958 abstract "Objective: The availability of new sequential media formulations for culture of human embryos to the blastocyst stage has made coculture less attractive. Concern about exposure of human embryos to animal proteins and recent FDA guidelines have virtually eliminated the use of monkey Vero cells in clinical laboratories in the U.S. Autologous endometrial coculture has been an alternative offered by some labs to aid patients with repeated failures and/or poor embryo quality, who may still potentially benefit from coculture methodology. In this study, we examine the feasibility of using a novel human endometrial cell line for coculture of embryos and also the Transwell chamber system to prevent direct contact between coculture cells and embryos. Design: Computer assisted image analysis along with differential labelling of ICM and trophectodermal cells. Materials and Methods: Frozen one-cell mouse embryos were purchased from Conception Technologies. Thawed zygotes were cultured overnight. The following day two cell embryos were pooled and randomly distributed between treatment groups. The treatment groups were as follows: 1) Control medium alone 2) endometrial coculture 3) Gardners G2.3 sequential medium. The control medium was alpha MEM supplemented with 10% Synthetic Serum Substitute (Irvine). For coculture, human endometrial cells were seeded into the outer well of a Transwell dish in control medium, the day before embryos were thawed. Two cell embryos were placed on the Transwell inserts at the outset of the experiment. Pore size of the Transwell inserts was 0.22 uM, allowing only exchange of fluids but no direct contact between the coculture cells and the embryos themselves. Control experiments were initially run to determine best cell concentration and to contrast with direct culture on cell monolayers. Embryo development and morphology in each culture regimen was monitored for 72 hrs. At termination of the experiment, cell nuclei were labelled with polynucleotide specific fluorochromes and cell counts were performed. All results were assessed using the student T-test and by chi square analysis. Results: The transwell coculture system produced blastocysts with overall higher cell numbers. Particularly striking was the overall percentage of developing embryos on day 5 that had reached a 100 or more cells (71%) versus only 13% in the G 2.3 series. The hatching rate was also higher within the coculture group. Tabled 1 Conclusion: Coculture with this novel human endometrial cell line appears to outperform the other treatments tested. The use of the Transwell system makes this coculture model particularly attractive for clinical use since direct contact between coculture cells and embryos can be avoided and no animal cells are involved. Preliminary trials with human embryos are underway." @default.
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• W2017462958 date "2003-09-01" @default.
• W2017462958 modified "2023-09-25" @default.
• W2017462958 title "Co-culture of embryos with novel human endometrial cell line in transwell dishes promotes embryo development without cell contact" @default.
• W2017462958 doi "https://doi.org/10.1016/s0015-0282(03)01912-5" @default.
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